摘要
目的检测线粒体转录因子A(mitochondrial transcription factor A,TFAM)在不同食管组织中的表达,探讨食管鳞癌中TFAM的表达与临床病理特征的关系,研究TFAM对食管鳞癌细胞增殖、克隆形成、迁移、侵袭和凋亡的影响及部分机制。方法SP免疫组化法检测食管低高级别上皮内瘤变、食管鳞癌及正常食管组织中TFAM蛋白的表达;以EC109、EC9076细胞为模型转染TFAM干扰质粒下调TFAM的表达,qRT-PCR测定转染效率,MTT、平板克隆、Transwell、流式细胞仪分别检测细胞增殖克隆形成、迁移、侵袭和凋亡能力;Western blot检测细胞内Bax、caspase3蛋白表达水平。结果TFAM在正常食管组织、食管低高级别上皮内瘤变、食管癌中的表达呈上升趋势(P<0.001),TFAM蛋白的表达与食管鳞癌临床分期、肌层浸润、淋巴结转移显著相关(P<0.05)。TFAM基因表达下调后,细胞的增殖、克隆形成、迁移及侵袭能力明显下降(P<0.05),凋亡率升高(P<0.001),Bax、caspase3蛋白表达量增加(P<0.001)。结论TFAM的表达异常可能是食管鳞癌发生发展的分子机制之一,下调TFAM可以抑制食管鳞癌细胞增殖、克隆形成、迁移和侵袭能力,并可能通过影响caspase3、Bax的表达来促进食管癌细胞的凋亡。
Objective This study aimed to detect the expression of mitochondrial transcription factor A(TFAM)in different esophageal tissues,explore the relationship between TFAM expression and clinicopathologicalcharacteristics in esophageal squamous cell carcinoma and study the effects of TFAM on the proliferation,colonyformation,migration,invasion and apoptosis of esophageal squamous carcinoma cells as well as some mechanisms.Methods SP immunohistochemical method was applied to detect the level of TFAM protein in low and high gradeintraepithelial neoplasia,esophageal squamous cell carcinoma and normal esophageal tissues. EC109 and EC9076 cells were transfected with TFAM interfering plasmids to down-regulate the TFAM expression,qRT-PCR to deter-mine transfection efficiency. MTT,plate cloning,Transwell,and flow cytometry were used to detect cell prolifera-tion,clone formation,migration,invasion,and apoptosis. Western blot was adopted to detect Bax and caspase3 protein expression levels.Results The expression of TFAM in normal esophageal tissue,low esophagus,high-grade intraepithelial neoplasia,and esophageal cancer showed an upward trend(P < 0.001). The expression ofTFAM protein was significantly correlated with clinical stage,myometrial invasion,and lymph node metastasis ofesophageal squamous cell carcinoma(P < 0.05). After down-regulation of TFAM gene expression,cell proliferation,colony formation,migration,and invasion abilities were significantly reduced(P < 0.05),and the apoptosis rateswere increased(P < 0.001),Bax and caspase3 protein levels were obviously increased(P < 0.001).Conclusion The abnormal expression of TFAM might be one of the molecular mechanisms of that contributed to the esophagealsquamous cell carcinoma. Down-regulation of TFAM could inhibit the proliferation,clone formation,migration andinvasion of esophageal squamous carcinoma cells,and might promote the apoptosis of esophageal carcinoma cellsby affecting the expression of caspase3 and Bax.
作者
王凯莉
陈卫刚
丁新
杨丹平
童娟娟
陈凯丽
张少聪
郑勇
WANG Kaili;CHEN Weigang;DING Xin;YANG Danping;TONG Juanjuan;CHEN Kaili;ZHANG Shaocong;ZHENG Yong(Shihezi University School of Medicine,Shihezi City 832000,China;不详)
出处
《实用医学杂志》
CAS
北大核心
2020年第9期1164-1171,共8页
The Journal of Practical Medicine
基金
国家自然科学基金资助项目(编号:81260362)。
